More than 1.3 billion people worldwide use tobacco products. Tobacco products are tightly regulated in most developed countries, and with these regulations, use has decreased. However, tobacco consumption is increasing globally due to increased consumption in developing nations. E-cigarettes are battery-powered nicotine delivery systems advertised as healthier than cigarettes that are gaining in popularity globally. By 2014, more than 466 brands existed with more than 7764 uniquely flavored nicotine solutions (e-liquids). While some countries have imposed restrictions sales of e-cigarettes, these devices are mostly unregulated. While the harmful effects of cigarette smoking in the orofacial region are recognized, little is known about the effects of aerosolized e-liquids. The goal of this applicatio is to unravel the effects of e-cigarette aerosol mixtures using a multi-system approach with robust in vitro and in vivo models to identify the most susceptible cell types and tissues, as well as the effects of these aerosol mixtures in the development, healing, and regeneration of craniofacial structures. Using cells from orofacial tissues as in vitro model, we will conduct biological, biochemical, and molecular interrogations to characterize the cytotoxic, proliferative, apoptotic, and inflammatory effects of aerosol mixtures. Additionally, we will use our novel Xenopus laevis model to elucidate the effects of e-cigarette aerosols in craniofacial development. These development studies will be complemented by our chronic exposure model, in which mice will be exposed to e-cigarette aerosols during pregnancy and the effects on their offspring's orofacial development studied. Our comprehensive approach will also investigate the effects of e-cigarette aerosols in our models of wound healing and regeneration, including common craniofacial surgical procedures like tooth extractions and dental implants, as well as more recent methods like bone tissue engineering. This approach will result in a detailed understanding of the effects of e- cigarette aerosol mixtures on the homeostasis, development, wound healing, and regeneration of craniofacial tissues.